人 TRF2 基因有效 siRNA 序列的筛选
Screening of Effectual siRNA-mediated TRF2
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摘要: 目的 设计合成干扰端粒重复序列结合因子2(telomeric repeatbinding factor 2,TRF2)基因表达的不同小型干扰RNA(siRNA)，筛选出能高效抑制TRF2 基因表达的siRNA。方法 根据人TRF2 mRNA的序列，设计合成3对不同的针对TRF2的siRNA、应用脂质体LipofectamineTM2000转染试剂将三对siRNA转染人乳腺癌MCF-7细胞，实时荧光定量RT－PCR技术检测TRF2 mRNA表达水平、Western印迹法检测TRF2蛋白表达以确定干扰效果。结果 设计合成的3对TRF2 siRNA中有2对siRNA可有效抑制TRF2 基因表达。结论 成功设计合成了针对TRF2基因的siRNA，并从中筛选出能特异且高效阻断TRF2表达的siRNA，为进一步应用RNAi技术抗TRF2基因进行肿瘤基因治疗研究奠定实验基础。Abstract: Objective To design and synthesize small interfering RNA（siRNA）targeting different regions of telomeric repeatbinding factor 2(TRF2) mRNA，and select the siRNA that can specifically and effectively suppresses TRF2 expression. Methods Three TRF2-specific double stranded siRNAs were designed and electroporated into MCF-7 cells by LipofectamineTM2000，The transcription level of TRF2 were analvzed by Real-time fluorescence quantitative RT-PCR and TRF2 protein was tested by western blotting. Results Two of the three customized TRF2-siRNAs could effectively inhibit the expression of TRF2. Conclusion TRF2-siRNAs can be successfully designed and synthesized，which can specifically and effectively suppresses the expression of TRF2 gene，the results of the study lay the foundation for further studying on gene therapy of tumor by RNA interfere（RNAi）.
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